QC Report


general
Report generated at2022-12-27 12:44:02
Titlelin-15B_OP184_L1larva_1_1
DescriptionENCSR799OPN
Pipeline versionv1.3.6
Pipeline typetf
GenomeWS245chr
Alignerbwa
Sequencing endedness{'rep1': {'paired_end': False}, 'rep2': {'paired_end': False}, 'rep3': {'paired_end': False}, 'ctl1': {'paired_end': False}}
Peak callerspp

Alignment quality metrics


Marking duplicates (filtered BAM)

rep1rep2rep3ctl1
Unpaired Reads309547099822919086432688840
Paired Reads0000
Unmapped Reads0000
Unpaired Duplicate Reads1248102821281293102770
Paired Duplicate Reads0000
Paired Optical Duplicate Reads0000
% Duplicate Reads4.0322.82624.25923.8221

Filtered out (samtools view -F 1804):


SAMstat (filtered/deduped BAM)

rep1rep2rep3ctl1
Total Reads297066097001718273502586070
Total Reads (QC-failed)0000
Duplicate Reads0000
Duplicate Reads (QC-failed)0000
Mapped Reads297066097001718273502586070
Mapped Reads (QC-failed)0000
% Mapped Reads100.0100.0100.0100.0
Paired Reads0000
Paired Reads (QC-failed)0000
Read10000
Read1 (QC-failed)0000
Read20000
Read2 (QC-failed)0000
Properly Paired Reads0000
Properly Paired Reads (QC-failed)0000
% Properly Paired Reads0.00.00.00.0
With itself0000
With itself (QC-failed)0000
Singletons0000
Singletons (QC-failed)0000
% Singleton0.00.00.00.0
Diff. Chroms0000
Diff. Chroms (QC-failed)0000

Filtered and duplicates removed


Sequence quality metrics (filtered/deduped BAM)

rep1
rep1
rep2
rep2
rep3
rep3

Open chromatin assays are known to have significant GC bias. Please take this into consideration as necessary.


Library complexity quality metrics


Library complexity (filtered non-mito BAM)

rep1rep2rep3ctl1
Total Fragments308146599422018735362630100
Distinct Fragments296086496647918094282565235
Positions with Two Read98183243705564049343
NRF = Distinct/Total0.9608620.9720980.9657820.975337
PBC1 = OneRead/Distinct0.964190.973220.9671820.978896
PBC2 = OneRead/TwoRead29.07668338.59651231.45303750.890643

Mitochondrial reads are filtered out by default. The non-redundant fraction (NRF) is the fraction of non-redundant mapped reads in a dataset; it is the ratio between the number of positions in the genome that uniquely mapped reads map to and the total number of uniquely mappable reads. The NRF should be > 0.8. The PBC1 is the ratio of genomic locations with EXACTLY one read pair over the genomic locations with AT LEAST one read pair. PBC1 is the primary measure, and the PBC1 should be close to 1. Provisionally 0-0.5 is severe bottlenecking, 0.5-0.8 is moderate bottlenecking, 0.8-0.9 is mild bottlenecking, and 0.9-1.0 is no bottlenecking. The PBC2 is the ratio of genomic locations with EXACTLY one read pair over the genomic locations with EXACTLY two read pairs. The PBC2 should be significantly greater than 1. See more details at the ENCODE portal standard for ChIP-Seq pipeline


NRF (non redundant fraction)
PBC1 (PCR Bottleneck coefficient 1)
PBC2 (PCR Bottleneck coefficient 2)
PBC1 is the primary measure. Provisionally


Replication quality metrics


IDR (Irreproducible Discovery Rate) plots

rep1_vs_rep2
rep1_vs_rep2
rep1_vs_rep3
rep1_vs_rep3
rep2_vs_rep3
rep2_vs_rep3
rep1-pr1_vs_rep1-pr2
rep1-pr1_vs_rep1-pr2
rep2-pr1_vs_rep2-pr2
rep2-pr1_vs_rep2-pr2
rep3-pr1_vs_rep3-pr2
rep3-pr1_vs_rep3-pr2
pooled-pr1_vs_pooled-pr2
pooled-pr1_vs_pooled-pr2

Reproducibility QC and peak detection statistics

overlapidr
Nt66451619
N16698412
N22059762
N385581414
Np24881118
N optimal66451619
N conservative66451619
Optimal Setrep2_vs_rep3rep2_vs_rep3
Conservative Setrep2_vs_rep3rep2_vs_rep3
Rescue Ratio2.67081993569131851.4481216457960644
Self Consistency Ratio4.1563865954346773.4320388349514563
Reproducibility Testfailborderline

Reproducibility QC


Number of raw peaks

rep1rep2rep3
Number of peaks114513063132314

Top 300000 raw peaks from spp with FDR 0.01

Peak calling statistics


Peak region size

rep1rep2rep3idr_optoverlap_opt
Min size75.060.085.072.072.0
25 percentile300.0240.0340.0290.0290.0
50 percentile (median)300.0240.0340.0290.0290.0
75 percentile300.0240.0340.0290.0290.0
Max size300.0240.0340.0290.0290.0
Mean299.79993013710595239.77581535046195339.26483876957354277.23038912909203286.8887885628292

rep1
rep1
rep2
rep2
rep3
rep3
idr_opt
idr_opt
overlap_opt
overlap_opt

Enrichment / Signal-to-noise ratio


Strand cross-correlation measures (trimmed/filtered SE BAM)

rep1rep2rep3
Number of Subsampled Reads30814659942201873536
Estimated Fragment Length110115135
Cross-correlation at Estimated Fragment Length0.5432546669373250.3094451302228280.439852166972061
Phantom Peak404040
Cross-correlation at Phantom Peak0.53807550.29363220.4205608
Argmin of Cross-correlation150015001500
Minimum of Cross-correlation0.52966760.27202510.4028237
NSC (Normalized Strand Cross-correlation coeff.)1.0256521.1375611.091922
RSC (Relative Strand Cross-correlation coeff.)1.6159911.7318372.087625


Performed on subsampled (15000000) reads mapped from FASTQs that are trimmed to 50. Such FASTQ trimming and subsampling reads are for cross-corrleation analysis only. Untrimmed FASTQs are used for all the other analyses.

NOTE1: For SE datasets, reads from replicates are randomly subsampled to 15000000.
NOTE2: For PE datasets, the first end (R1) of each read-pair is selected and trimmed to 50 the reads are then randomly subsampled to 15000000.


rep1
rep1
rep2
rep2
rep3
rep3

Jensen-Shannon distance (filtered/deduped BAM)

rep1rep2rep3
AUC0.369300574245421230.31507149067289270.32450650626550687
Synthetic AUC0.49350164957386250.488616671478502740.4917156096925494
X-intercept0.040242805306927690.06529930760990770.04772724099204679
Synthetic X-intercept7.841135688659982e-2032.7531429241172766e-652.9160889261399934e-124
Elbow Point0.492949430189484670.54242410339372880.5946834973379586
Synthetic Elbow Point0.50263145310945590.49903571873440330.5127055171540771
JS Distance0.036783852238579560.107745064114037530.09937735400807503
Synthetic JS Distance0.17389206236061290.229527515185407120.23654053595318375
% Genome Enriched38.6507668953355935.2372897246597436.56721330442224
Diff. Enrichment7.66855522580071117.37268518706810417.564356347039467
CHANCE Divergence0.065214544129513440.147998131662668870.14962250750703707

Peak enrichment


Fraction of reads in peaks (FRiP)

FRiP for spp raw peaks

rep1rep2rep3rep1-pr1rep2-pr1rep3-pr1rep1-pr2rep2-pr2rep3-pr2pooledpooled-pr1pooled-pr2
Fraction of Reads in Peaks0.075348912362909250.218369368784258420.245370071414890420.23281627651767620.141395314313755020.267777929789038750.230688129910524930.14592336621251610.26860426300380330.43558152553724180.071335645388683960.07943584165536008

FRiP for overlap peaks

rep1_vs_rep2rep1_vs_rep3rep2_vs_rep3rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.0402884729908511150.0404497066327879560.0622091401444549440.048717456726787980.052707323686079730.10804498317235340.04163208667365808

FRiP for IDR peaks

rep1_vs_rep2rep1_vs_rep3rep2_vs_rep3rep1-pr1_vs_rep1-pr2rep2-pr1_vs_rep2-pr2rep3-pr1_vs_rep3-pr2pooled-pr1_vs_pooled-pr2
Fraction of Reads in Peaks0.0223929256919220380.0219707709412594630.0340565326757312350.0099028498717456720.0336602348206268550.050768599337838950.02785579887195396

For spp raw peaks:


For overlap/IDR peaks: